Daunorubicin efflux against a concentration gradient in non-P-glycoprotein multidrug-resistant lung-cancer cells

H. S. Mulder, J. Lankelma, H. Dekker, H. J. Broxterman, H. M. Pinedo

Research output: Contribution to journalArticle

Abstract

Multidrug-resistant, human non-small-cell lung carcinoma SW-1573/2R120 (2R120) cells, not containing the drug efflux pump P-glycoprotein (Pgp), have reduced initial daunorubicin (DN) accumulation rates and decreased cellular steady-state drug concentrations. Previously we found indications of the presence of a plasma membrane 'vacuum cleaner', pumping DN directly from the membrane, and reported evidence of active DN pumping using digitonin. Further evidence of active DN pumping is now provided via a different methodology and the active drug pump flux is estimated. Cells were exposed to a flowing medium containing the cytotoxic agent DN. After reaching a steady state, in which net DN uptake equals net DN efflux, high concentration pulses of vincristine (VCR) were injected into the flowing medium. A rapid increase in cellular DN content was observed, while only a minimal effect was seen in SW-1573 wild-type cells. After passage of the VCR pulse, the extra accumulated DN was effluxed against a concentration gradient. Upon increasing the VCR concentration, a maximum pump inhibition was reached which was similar to the effect of cellular energy depletion. Similar effects were observed for Pgp-containing SW-1573/2R160 (2R160) cells as well as non-Pgp MDR human small-cell lung carcinoma GLC4/ADR cells. With increasing extracellular DN concentrations, saturation of the VCR-induced DN influx was observed (DN medium concentration 2.5 μM at 1/2 V(max). At an extracellular DN concentration of 5 μM, higher concentrations of VCR were needed to reach the maximum effect in 2R120 cells than at 0.5 μM DN. This is an indication of competitive interaction between DN and VCR for the putative DN efflux system. In summary, we found indications of inhibition of active DN efflux by VCR and DN efflux against a concentration gradient in non-Pgp MDR 2R120 and GLC4/ADR cells. These features are consistent with the presence of a multidrug transporter, different from Pgp, in the plasma membrane of these cells.

Original languageEnglish (US)
Pages (from-to)275-281
Number of pages7
JournalInternational Journal of Cancer
Volume59
Issue number2
DOIs
StatePublished - 1994
Externally publishedYes

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Daunorubicin
Lung Neoplasms
Glycoproteins
Vincristine
P-Glycoprotein
Cell Membrane
Pharmaceutical Preparations
Digitonin
Cytotoxins
Small Cell Lung Carcinoma

ASJC Scopus subject areas

  • Cancer Research
  • Oncology

Cite this

Daunorubicin efflux against a concentration gradient in non-P-glycoprotein multidrug-resistant lung-cancer cells. / Mulder, H. S.; Lankelma, J.; Dekker, H.; Broxterman, H. J.; Pinedo, H. M.

In: International Journal of Cancer, Vol. 59, No. 2, 1994, p. 275-281.

Research output: Contribution to journalArticle

Mulder, H. S. ; Lankelma, J. ; Dekker, H. ; Broxterman, H. J. ; Pinedo, H. M. / Daunorubicin efflux against a concentration gradient in non-P-glycoprotein multidrug-resistant lung-cancer cells. In: International Journal of Cancer. 1994 ; Vol. 59, No. 2. pp. 275-281.
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abstract = "Multidrug-resistant, human non-small-cell lung carcinoma SW-1573/2R120 (2R120) cells, not containing the drug efflux pump P-glycoprotein (Pgp), have reduced initial daunorubicin (DN) accumulation rates and decreased cellular steady-state drug concentrations. Previously we found indications of the presence of a plasma membrane 'vacuum cleaner', pumping DN directly from the membrane, and reported evidence of active DN pumping using digitonin. Further evidence of active DN pumping is now provided via a different methodology and the active drug pump flux is estimated. Cells were exposed to a flowing medium containing the cytotoxic agent DN. After reaching a steady state, in which net DN uptake equals net DN efflux, high concentration pulses of vincristine (VCR) were injected into the flowing medium. A rapid increase in cellular DN content was observed, while only a minimal effect was seen in SW-1573 wild-type cells. After passage of the VCR pulse, the extra accumulated DN was effluxed against a concentration gradient. Upon increasing the VCR concentration, a maximum pump inhibition was reached which was similar to the effect of cellular energy depletion. Similar effects were observed for Pgp-containing SW-1573/2R160 (2R160) cells as well as non-Pgp MDR human small-cell lung carcinoma GLC4/ADR cells. With increasing extracellular DN concentrations, saturation of the VCR-induced DN influx was observed (DN medium concentration 2.5 μM at 1/2 V(max). At an extracellular DN concentration of 5 μM, higher concentrations of VCR were needed to reach the maximum effect in 2R120 cells than at 0.5 μM DN. This is an indication of competitive interaction between DN and VCR for the putative DN efflux system. In summary, we found indications of inhibition of active DN efflux by VCR and DN efflux against a concentration gradient in non-Pgp MDR 2R120 and GLC4/ADR cells. These features are consistent with the presence of a multidrug transporter, different from Pgp, in the plasma membrane of these cells.",
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AU - Broxterman, H. J.

AU - Pinedo, H. M.

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AB - Multidrug-resistant, human non-small-cell lung carcinoma SW-1573/2R120 (2R120) cells, not containing the drug efflux pump P-glycoprotein (Pgp), have reduced initial daunorubicin (DN) accumulation rates and decreased cellular steady-state drug concentrations. Previously we found indications of the presence of a plasma membrane 'vacuum cleaner', pumping DN directly from the membrane, and reported evidence of active DN pumping using digitonin. Further evidence of active DN pumping is now provided via a different methodology and the active drug pump flux is estimated. Cells were exposed to a flowing medium containing the cytotoxic agent DN. After reaching a steady state, in which net DN uptake equals net DN efflux, high concentration pulses of vincristine (VCR) were injected into the flowing medium. A rapid increase in cellular DN content was observed, while only a minimal effect was seen in SW-1573 wild-type cells. After passage of the VCR pulse, the extra accumulated DN was effluxed against a concentration gradient. Upon increasing the VCR concentration, a maximum pump inhibition was reached which was similar to the effect of cellular energy depletion. Similar effects were observed for Pgp-containing SW-1573/2R160 (2R160) cells as well as non-Pgp MDR human small-cell lung carcinoma GLC4/ADR cells. With increasing extracellular DN concentrations, saturation of the VCR-induced DN influx was observed (DN medium concentration 2.5 μM at 1/2 V(max). At an extracellular DN concentration of 5 μM, higher concentrations of VCR were needed to reach the maximum effect in 2R120 cells than at 0.5 μM DN. This is an indication of competitive interaction between DN and VCR for the putative DN efflux system. In summary, we found indications of inhibition of active DN efflux by VCR and DN efflux against a concentration gradient in non-Pgp MDR 2R120 and GLC4/ADR cells. These features are consistent with the presence of a multidrug transporter, different from Pgp, in the plasma membrane of these cells.

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